Project 1 - Determining the contributions of the gut microbiota to the physiologi

摘要 项目1将使用转译研究管道进行人体肠道微生物区系导向的诊断和 我们开发的治疗方法是基于携带从三种类型的 与肥胖和/或相关代谢异常不一致的成年双胞胎[瘦身代谢健康肥胖 代谢不良(LnMH/ObMUN)、LnMH/ObMH、ObMH/ObMUN]。它的目标是(I)建立肠道的因果关系 微生物区系在肥胖和相关的代谢表型中，(Ii)获得关于 饮食和产生这些表型的肠道微生物区系成员之间的相互作用，(Iii)进行测试 控制饮食和细菌分类群对ObMun微生物区系传播的表型的影响。项目1 有四个目标。(1)确定从双胞胎对每对结束时收集的微生物区系的影响 居家饮食周期对受体成年灵知生菌小鼠体成分/代谢表型的影响 与采集捐献者微生物区系时消耗的饮食同源或双胞胎的其他饮食 将已进食(交叉节食组)。双胞胎对内、双胞胎对间比较和交叉不协调 将执行配对类型比较。(2)共居灵知生菌小鼠，含有完整的未培养微生物区系 从不一致的配对中确定(I)来自LnMH供体的微生物区系是否阻止或改善了 带有ObMH或ObMUN同卵双胞胎的小鼠的肥胖及肥胖相关代谢功能障碍 微生物区系，以及预防/改善与来自微生物区系的细菌分类群入侵之间的关系 一个笼子到另一个笼子，后来进行了涉及ObMH和ObMUN小鼠的可比较的共居实验。(3) 确定从目标2中表征的微生物区系样品制备的细菌培养样本是否也转移 不一致的供体表型；进行共居实验以鉴定侵入性培养的 在不同的饮食环境中与表型拯救相关的分类群。(4)执行测试矩阵，其中文化 从具有代表性的ObMUN或ObMH同卵双胞胎中收集的样品被引入到不同的诺生菌小鼠组中 单独或加铅喂养具有代表性的美国饮食，高饱和脂肪，低水果蔬菜 益生菌联合体(从AIMS 2，3鉴定的入侵分类群)，或铅益生菌(从体外筛选鉴定)， 或两者的组合(合生体铅)。治疗性铅将在以下时间给予 在初始定植后2周用培养物收集定植(预防组)或定植后2周(治疗组)定植。 代谢分析将使用Core A进行，多组学数据集将使用现有工具和 用于为项目3和核心B生成新的分析策略。

SUMMARY Project 1 will use a translational research pipeline for human gut microbiota-directed diagnostics and therapeutics we developed based on gnotobiotic mice harboring microbiota transplanted from three types of adult twin pairs discordant for obesity and/or its associated metabolic abnormalities [LeanMetabolicallyHealthy-Obese Metabolically Unhealthy (LnMH/ObMUN), LnMH/ObMH, ObMH/ObMUN]. Its goal is to (i) establish a causal role of the gut microbiota in obesity and associated metabolic phenotypes, (ii) obtain mechanistic insights about the interactions between diet and members of the gut microbiota that produce these phenotypes, (iii) conduct tests of the effects of manipulating diet and bacterial taxa on phenotypes transmitted by ObMUN microbiota. Project 1 has 4 aims. (1) Determine the effects of microbiota, collected from twin pairs at the end of the each of their two in-home diet periods, on body composition/metabolic phenotypes of recipient adult gnotobiotic mice given a diet homologous to that consumed at the time the donor's microbiota was collected or the other diet that twins will have consumed (cross-over diet group). Within-pair, between-twin pair comparisons and across discordant pair type comparisons will be performed. (2) Co-house gnotobiotic mice harboring intact uncultured microbiota from discordant pairs to determine if (i) microbiota from the LnMH donor prevents or ameliorates development of obesity- and obesity-associated metabolic dysfunction in mice colonized with the ObMH or ObMUN co-twin's microbiota, and how prevention/amelioration correlates with invasion of bacterial taxa from the microbiota of one cagemate to the other, and later comparable co-housing experiments involving ObMH and ObMUN mice. (3) Determine if bacterial culture collections prepared from microbiota samples characterized in Aim 2 also transfer discordant donor phenotypes to gnotobiotic mice; perform co-housing experiments to identify invasive cultured taxa associated with phenotypic rescue in different diet contexts. (4) Execute a testing matrix in which a culture collection from a representative ObMUN or ObMH co-twin are introduced to separate groups of gnotobiotic mice fed a representative USA diet high in saturated fats and low in fruits and vegetables, alone or with a lead probiotic consortium (invasive taxa identified from aims 2, 3), or a lead prebiotic (identified from in vitro screen), or a combination of the two (synbiotic lead). The therapeutic lead will be administered at the time of colonization with the culture collection (prevention arm) or 2 weeks after initial colonization (treatment arm). Metabolic profiling will be performed with Core A. Multi-omics datasets will be analyzed with existing tools and used to generate new analysis strategies with Project 3 and Core B.